ES2435085T3 - Protection of plants against their pathogens - Google Patents

Abstract

Use of walls or bark of yeast (s) as active material, for the treatment or protection of plants against diseases caused by pathogens or for the induction or stimulation in a plant of natural defenses against pathogens.

Description

Protection of plants against their pathogens

Field of the Invention

The present invention relates to methods for protecting plants against various pathogens such as fungi, viruses and bacteria. The invention is therefore usable, alone or in alternation and / or in combination with other means of plant protection, and is adapted to the treatment of multiple plant varieties.

Among the pathogens, fungi, responsible for fungal or cryptogamic diseases, have the greatest economic impact. Each plant species is sensitive to one or several major diseases, likely to strongly reduce its vigor, its growth and finally the quantity and / or quality of the crop.

Various parameters influence the development of diseases such as soil conditions and fertilization, varietal sensitivity, mode of behavior (rotation, soil work, number of plants or seedlings per hectare, pruning system, etc.), or over All weather conditions. But acting on some of these parameters is generally not enough to sufficiently limit the damage caused by diseases. Also, to prevent, the technician who wishes to optimize and ensure its performance, will treat your crop in a timely manner with a phytosanitary product, often preventive. Most commonly the products used are chemical products, most of them very effective, but may present health risks to the people who handle them and generate waste on the treated products, in the soil and water. In addition, the repeated use of certain fungicidal active substances that act on the same metabolic site selects strains resistant to these fungicides.

To try to remedy it, it is necessary to limit the number of annual uses of chemical products of the same family, to alternate chemical families with different modes of action, and to use any other unfavorable means to the pathogen.

In this context, there is therefore a real and important need for alternative solutions against plant diseases. Ideally, these solutions should act differently from existing chemical fungicides, not generate chemical residues in crops and the environment, and be safer and healthier for operators. Such treatments could be used, alone or in alternation and / or in combination with current chemical treatments or any other treatment, to prevent the occurrence or limit the development of these pathogens, and their resistant strains, on plants, and limit the risks to The man and the environment.

Summary of the Invention

The present invention provides new methods for the treatment or protection of plants against pathogens. More particularly, the invention resides in showing that, unexpectedly, the walls

o Yeast barks have the ability to effectively protect plants against infection by pathogens. This effect is obtained by simple contact of the walls or bark with the plant, by means of a spray for example.

The results obtained show that the treatment by means of walls or bark of yeasts confers protection not only to the organs or parts of treated plants (direct action at the point of contact), but also to the organs that appear later. Since the walls or crusts cannot penetrate the plant because they are vehicularized by the sap, it is not a system. On the contrary, this result suggests an inducing effect of natural defense mechanisms of the plant, and therefore allows for a long duration of protection, equal to or greater than one month, and a versatile action of the compositions and methods of the invention. .

An object of the invention therefore resides in the use of walls or bark of yeast (s) as active material, for the treatment or protection of plants against diseases caused or caused by pathogens, mainly fungal, bacterial or viral. .

Another object of the invention is the use of walls or bark of yeast (s) for induction or stimulation in a plant of natural defenses against pathogens.

The invention also relates to the use for the treatment or protection of plants against diseases caused or caused by pathogens, mainly fungal, bacterial or viral, comprising the application on the plant or one of its parts of walls or yeast bark (s).

The invention also aims at the use for induction or stimulation in a plant of natural defenses (against pathogens), comprising the application on the plant or one of its parts of walls or yeast bark (s).

As will be described in more detail below in the text, the invention is applicable to all types of plants, and mainly to grasses and dicotyledons, to annual, bi-annual and perennial plants, to vegetables, to cereals of which the wheat, barley and rice, corn, sorghum, millet, oilseeds, protein crops, potatoes, beets, sugar cane, tobacco, woody plants, trees, fruit or not, vineyards To decorative vegetables, etc. In addition, the pathogen can be of a diverse nature, such as a fungus, a bacterium, a virus, a mycoplasma, a spiroplasma or a viroid.

The walls or bark of yeast (s) used in the invention can come from any species of yeast, in particular yeasts of the genus Saccharomyces, mainly S.cerevisiae, and can be obtained or prepared according to techniques known to those skilled in the art, which they will be described below in the text, mainly by autolysis, separation, concentration, etc.

Also described is a composition (phytopharmaceutical) comprising walls or bark of yeast, being able to be administered or applied to a plant, or to put it in contact with it, or some of its organs only, whatever they are, chosen mainly among the leaves, flowers, fruits, stem, trunk or roots. Such a composition can be described as a phytopharmaceutical preparation.

Said composition may be in liquid form, concentrated or not, powder, wettable or not, granulated, dispersible or other, or in any other way adapted to bring into contact walls or bark of yeasts with the plant or organs of the plant to which It aims the treatment, for example, spraying after dilution, suspension or other, in water or in another vehicle, on the aerial parts of the plant, contribution to the soil or by nutrient solution, at the level of the roots of the plant, etc.

In preferred embodiments, the compositions of the invention may further comprise formulation agents, dispersants, stabilizers, surfactants, etc.

A composition (phytopharmaceutical) comprising walls or bark of yeast in combination with a fungicidal, antiviral or antibacterial agent is also described, with a view to simultaneous, separate or time-spaced application.

Furthermore, another objective of the invention lies in the use to fight against diseases caused by pathogens in plants, which includes the application on a plant of bark or yeast walls (s), possibly in alternation or in combination with another active treatment against said pathogen.

Another objective of the invention lies in the use to prevent or curb the development of pathogens resistant to a family of active substances, characterized in that the plant is treated with bark or yeast walls (s), with a view to reducing the pressure selection of the strains resistant to said family of active substances, or why the treatment (s) of the plant with a substance of said family of active substances alternates or combines with a treatment (s) of said plant with bark or walls of yeast (s), or a composition comprising bark or walls of yeast (s).

The use of a composition comprising bark or walls of yeast (s) is also described, in order to increase the effectiveness of the entire phytosanitary protection, by decreasing the level of infection and / or reducing the emission of inoculum.

It is also described the use of a composition comprising bark or walls of yeast (s), to obtain a partial or complete long-term phytosanitary protection, for example of at least a month and a half.

The invention describes a method for limiting the amount of residues of agrochemicals in or on consumable products, in soils and waters during the treatment of crops or a plant, the method comprising a treatment (s) of said crop or plant by crusts or walls of yeast (s), or a composition comprising crusts or walls of yeast (s).

The invention describes the use of bark or walls of yeast (s), or a composition comprising bark or walls of yeast (s), for the prevention or treatment of diseases in organic or organic farming.

Detailed description of the invention

As indicated above, the invention relates to the use of products to fight, on a preventive or curative basis, against plant pathogens. The invention relies mainly on revealing the advantageous and unexpected properties of walls and bark of yeast, and proposes its use in the fight against diseases caused by pathogens in plants.

The present invention derives, mainly, from the discovery that the bark and / or yeast walls are capable of protecting the organs or parts of the plant treated directly by the compositions of the invention (i.e. that they exert a direct action on the site of contact), and also the organs or parts of the plant that appear after treatment. Since the walls or barks cannot penetrate the plant, because they are vehiculized by the sap, it is not a system. On the contrary, this propagation of protection to the whole plant and to the newly formed organs suggests a mechanism of induction or stimulation of natural defenses, such as those already known with b-aminobutyric acid, 2,6-dichloroisonicotinic acid, acibenzolar- s-methyl, or certain algae extracts (patents or patent applications No. FR2 868 253; WO 03/092384; WO 97/14310 and WO 99/53761) or also with oligo 1 (1-3) glucans (WO 99 / 03346) or chitin or chitosan (Ozereskovskaya OL et al. (Russian Journal of Plant Physiology, 43 (5), 1996, 648-655) and allows for a long term of protection, equal to or greater than one month, and a polyvalent action of the compositions of the invention.

In addition, the walls or bark of yeast according to the invention that do not have a priori direct effect on the pathogens, are less likely to cause resistance phenomena.

The results presented in the invention are particularly surprising since they show that it is not necessary to use purified molecules, but rather that the entire yeast walls or barks are active without their components having been previously isolated or separated. In addition, it is not necessary to proceed with subsequent chemical modifications of the yeast crusts or walls.

The invention therefore relates to the use of walls or bark of yeast (s) as an active material, for the protection of plants against diseases caused by pathogens.

The invention also relates to the use of walls or bark of yeast (s) as an active material, for the induction or stimulation in a plant of natural defenses against pathogens.

The invention also relates to the use for induction or stimulation in a plant of natural defenses against pathogens, which comprises the application on the plant or one of its parts of walls or yeast bark (s).

The walls and yeast barks used in the present invention can be produced from different types of yeasts, possibly in mixture (s). The yeast is preferably a baker's yeast. A baker's yeast is a yeast that belongs to the genus Saccharomyces, essentially produced by multiplication

or aerobic culture as shown in chapter 6 “Backer’s yeast production” of the reference work “Yeast technology”. The yeast can also be a brewer's yeast, an oenological yeast or a distillery yeast. Other types of yeast are usable within the framework of the present invention such as for example yeasts of the genus Kluyveromyces spp, Pichia spp, Metschnikowia spp or Candida spp.

A yeast is a cell, schematically composed of a wrapper and a content. The wrapper is

called "bark" or "wall."

Industrial products called "walls" or "barks" of yeasts can be produced in different ways, from different types of yeast (s), possibly in mixture (s), following protocols that are known to those skilled in the art.

In a particular embodiment, the yeast crusts or walls (s) are susceptible to being produced by lysis (autolysis or heterolysis) of yeast cells, for example Saccharomyces cerevisiae, followed by a separation of the soluble and insoluble parts, for example by physical means, such as centrifugation, and recovery of the insoluble part. The insoluble part is thus typically recovered by elimination of the soluble part.

by centrifugation. The insoluble part is called "yeast barks" or "yeast walls." The soluble part resulting from this procedure, of light color and low turbidity, is called "yeast extract".

Yeast autolysis is a hydrolysis of the cellular content of the yeast, by its own enzymes. It is typically obtained by placing a yeast suspension (s) under certain physical conditions of medium and / or in contact with activators that cause the death of yeasts and the release of their enzymes in the cell body. Hydrolysis of cellular content produces soluble compounds. Separated and recovered this soluble fraction constitutes the

"yeast extract". The recovered insoluble fraction derived from said separation constitutes the product called "yeast barks" or "yeast walls". This product includes the cytoskeleton of the

yeasts, and membranes and components not solubilized by autolysis or heterolysis. The insoluble fraction is generally recovered in the form of an aqueous suspension of yeast barks, whose concentration in dry matter is typically 10 to 15% (weight / volume). The crusts correspond to approximately 25 to 45% of the dry weight of whole yeast cells, on average approximately 35%.

Barks or walls that have undergone additional chemical treatments, extraction or functionalization can be used, for example, as a delipidation. In a preferred embodiment of the invention, unmodified yeast crusts or walls are used, that is to say mainly not subjected to such treatments.

For the realization of the present invention, the walls or bark of yeast (s) or any composition comprising walls or bark of yeast (s) that come from the same type of yeast or of several different types or species can be used.

Such products are also commercially available, such as mainly Springcell 8001 0 PW from Biospringer SA (F-94 Maison-Alfort) or Pronady de Prodesa (Bra.Valinhos).

For the realization of the invention, any preparation comprising walls or barks of yeasts, more or less dehydrated, can be used as raw material. In a preferred embodiment, an aqueous suspension of bark or yeast walls is used, preferably concentrated to less than 20% dry matter, preferably less than 17% or even 14% dry matter. In another preferred embodiment, preparations derived from drying the liquid suspension are used, for example by atomization, and preferably containing more than 80% dry matter, preferably more than 85, 90, 93 or 95%.

In a particular embodiment of the invention, more or less concentrated phytosanitary (or phytopharmaceutical) compositions (or preparations) are prepared, intended to be mixed for application with a liquid or solid vehicle, and comprising, as active material, cortices and / or yeast walls (s) as defined above. Professional agriculture often uses concentrated phytosanitary preparations that are diluted in water for spraying, or mixed with a fertilizer or fertilizer for soil supply. In a particular way, the composition of the invention is therefore a concentrated phytosanitary composition, of dry or liquid formulation.

According to another particular embodiment of the invention, ready-to-use compositions are prepared, which may be in liquid or solid form. In a preparation ready for use, the active material (comprising the crusts or walls of yeast (s)) is already mixed with a vehicle adapted for use in plants, such as a liquid to refill a spray type spray , a fertilizer, a greenhouse cultivation substrate, etc.

Such preparations or compositions may comprise, in addition to the active material, any adapted formulation agent.

Thus, any composition, mainly of the phytopharmaceutical or phytosanitary type, comprising yeast walls or barks can be used according to the invention.

According to a first variant embodiment, the composition is a dry form composition, for example powder or granulate.

According to another variant embodiment, the composition is a composition in liquid form, preferably in aqueous liquid form. It can be mainly a suspension, a gel, a cream, a paste, etc.

In a preferred embodiment, the compositions further comprise one or more formulation agents. In general, the compositions according to the invention comprise approximately 0.1% to 99.9% (by weight) of active material and one or more formulation agents, solid or liquid.

The formulation agent (s) may be constituted by any compound or any inert material that makes it possible to facilitate, facilitate, or optimize the transport, storage, handling, and / or application of the active material on the plant or parts of it. Such agents are adapted to the desired objective: preservation of the active agents, suspension maintenance of the walls or barks or other active substances during storage or during use in the preparation of the treatment porridge, antifoam, anti-dust, adhesion to the plant, and others. This or these agents can be solid (s), liquid (s), alone or mixed.

The formulation agent (s), and especially those adapted for spraying, can be chosen primarily from all surfactants, dispersants, preservatives, humectants, emulsifiers, adhesion agents, pH buffer, etc., alone or in admixture.

In a particular embodiment, the composition further comprises another active agent, preferably a fungicidal, antibacterial or antiviral agent. The fungicidal agent can be chosen, for example, from organic agrochemical fungicides, or inorganic mineral fungicides based on sulfur and / or copper, for example.

Examples of currently available organic agrochemical fungicides are mainly chloronitriles among which chlorothalonyl, carbamates among which dithiocarbamates such as mancozeb, phthalimides among which captane, sulfonamides, guanidines, quinones, quinoleins, thiadiazines, anilides, hydroxyanilides, and phenylamides, imidazolinones, oxazolidinediones, strobilurines, cyanoimidazoles, fluazinam, diconap, sitiofam, dicarboximides, fludioxonil, organophosphates, propamocarb HCl, diphenylamine, pyridylamines, sterile biosynthesis inhibitors (IBS), among which pyrimimines, trimidimines, tripidimines, hydroxylimines, tridimines, tripyrimines Spiroxamine, morpholines and piperidines, fenhexamide, himexazole, zoxamide, dietofencarb, benzimidazoles, pencicuron, quinoxifene, iprovalicarb, cymoxanil, dimetomorph, phosphonates, triazines, etc.

The invention can be used in alternation, association or combination with one or more eliciters of defense compounds in plants, such as, for example, b-aminobutyric acid, 2,6-dichloroisonicotinic acid, acibenzolar-smethyl, or certain algae extracts (patents or Patent applications No. FR2 868 253; WO 03/092384; WO 97/14310 and WO 99/53761). Examples of such compounds are mainly laminarin and ulvans.

The products or compositions used according to the invention can be applied in different ways and according to different protocols or treatment programs.

In a preferred embodiment, the products or compositions are applied by spraying, mainly foliar or soil spraying.

In a variant, it is possible to apply the products or compositions in the form of a mixture with fertilizers, cultivation support, in the irrigation water, or others. The composition can thus be administered in the roots by spraying in the soil, mechanical incorporation, mixed with fertilizers, fertilizers, in premix, or others.

Thus the invention describes any composition, mainly of the phytopharmaceutical (or phytosanitary) type or ready for use, which comprises walls or bark of yeast as active material. Such compositions advantageously comprise one or excipients adapted to an application on vegetables, such as in the form of a spray, a spray, or a powder, mainly for domestic or gardening use. Such compositions may further comprise one or more supplementary active agents, such as for example a fungicidal, antiviral, or one or fertilizer agent, with a view to their simultaneous, separate or sequential application on vegetables. Other compositions ready for use, for example mixed with a fertilizer to contribute to the soil, or a culture support are usable.

The products or compositions used according to the invention can be applied on the whole plant or on one or only parts thereof, such as for example the leaves, stems, flowers, fruits, trunk and / or roots. They can also be used on plant propagation material, such as seeds, grains or plants, in briquettes or not. Taking into account its proposed mode of action, the products of the invention should allow the vegetables to be effectively protected against pathogens for an important period of time, and may exceed one month. Of course, a repeated application can be provided, at intervals to be defined by the user.

The amount applied is defined by the person skilled in the art, mainly depending on the pathogen to be treated, the type of vegetable, combinations used, etc. The amount applied is preferably sufficient to protect the plant against a pathogen, or to limit or suppress the development and effects of the pathogen present. This quantity can be determined, for example, with field tests.

According to the invention, the composition is applied or used at an effective use dose greater than 1 mg / l of bark or yeast walls when the product is sprayed to the limit of runoff, or greater than 1 g / ha in the case of spraying with low water volume. Preferably, the effective dose is between 1 and 1000 mg / l of cortices or yeast walls when the product is sprayed to the limit of runoff, or also between 1 and 1000 g / ha in other cases.

In a particular embodiment, the composition is applied or used at a dose of effective use between 1 and 250 mg / l of bark or yeast walls, preferably between 2.5 mg and 25 mg / l, when the product it is applied by spraying to the runoff limit, or also between 1 and 250 g / ha, preferably between 2.5 and 25 g / ha in other cases, such as in the case of spraying with low water volume. Regardless of the dose used, the composition can be produced, transported and / or sold in various concentrations. Thus, when the preparation is in dry form, it can contain, for example, 96% by weight of walls or bark of yeasts. A liquid preparation may be in the form of a suspension comprising, for example, 13% of walls or barks of yeast in dry matter. The preparation may also be ready for use, that is, for example, to comprise walls or bark of yeasts at a concentration of approximately 25 mg / l. It is understood that the concentration in active matter in the preparations of the invention or during its application can be adapted by the person skilled in the art, and that the doses higher than those mentioned above can be used.

In addition, as indicated above, the products and compositions of the invention can be used in alternation and / or in combination with one or more other treatments.

A particular objective of the invention lies in a use to combat fungal diseases in plants, which comprises the application on a bark plant or yeast walls (s), in alternation or in combination with an antifungal treatment.

Another objective of the invention lies in a use to fight against diseases caused by bacteria in plants, which comprises the application on a bark plant or yeast walls (s), in alternation

or in combination with an antibacterial treatment.

Another object of the invention lies in a use for preventing or curbing the development of fungal strains resistant to a family of fungicidal agents, characterized in that the plant is treated with bark or yeast walls (s), or a composition comprising bark or yeast walls (s), with a view to reducing the selection pressure of the strains resistant to said family of fungicidal agents, or because the treatment (s) of the plant with a substance of said family of fungicidal agents is (are) alternated (s) or combined (s) with a treatment (s) of said plant with bark or walls of yeast (s), or a composition comprising bark or walls of yeast (s).

Another object of the invention lies in a use to prevent or slow the development of strains of bacteria resistant to a family of antibacterial agents, characterized in that the plant is treated with bark or yeast walls (s), or a composition comprising barks or yeast walls (s), with a view to reducing the selection pressure of the strains resistant to said family of antibacterial agents, or because the treatment (s) of the plant with said family and antibacterial agent is (are ) alternated (s) or combined (s) with a treatment (s) of said plant with bark or walls of yeast (s), or a composition comprising bark or walls of yeast (s).

The invention is applicable to the treatment of all types of plants, in the field, orchard, forest, greenhouse or indoor or garden plants. The invention is thus applicable mainly to grasses and dicotyledons, to annual, bi-annual and perennial plants, to leguminous plants, to cereals such as wheat, barley and rice, to corn, sorghum, millet, oilseeds, proteaginous plants , potatoes, beets, sugar cane, tobacco, woody plants, trees, fruit or not, vineyards, decorative vegetables, etc.

In a first particular embodiment, the plant is a fruit tree, for example a fruit tree with seeds in particular chosen among apple, pear and citrus.

In another particular embodiment, the plant is chosen among the vineyard, cereals mainly wheat, rapeseed, beets, potatoes, beans, tomatoes, cucumbers, lettuce or strawberries.

It is understood that the invention will not be limited to a particular type of plant, but can be used on all vegetables.

The invention can be applied to the fight against all types of pathogens, and mainly fungi, viruses, bacteria, mycoplasmas, spiroplasmas or viroids. By way of particular examples of pathogens, fungi of the genera Alternaria spp, for example A. solani, Ascochyta spp, for example, A. fabae or A. pinodella, Botrytis spp, for example, B. cinerea, Bremia spp, for example B. lactucae, Cercospora spp for example C. beticola, Cladosporium spp for example C. allii-cepae, Colletotrichum spp for example C. graminícola, Erysiphe spp for example E. graminis, Fusarium spp for example F. oxysporum and F. roseum, Glocosporium spp for example G. fructigenum, Guignardia spp for example G. bidwellii, Helminthosporium spp for example H. triticirepentis, Marssonina spp for example M. rosae, Monilia spp for example M. fructigena, Mycosphaerella spp for example M. brassicicola, Penicilium spp for example P. expansum or P. digitatum, Peronospora spp for example P. parasitica, Pezicula spp Phragmidium spp for example P. rubi-idaei Phytophtora spp of which P. infestans, Plasmopara spp of which P. viticola , Podosphaera spp for example P. leucotricha, Pseudocercosporella spp of which P. brassicae, Pseudoperonospora spp for example P. cubensis, Pseudopeziza spp for example P. medicaginis, Puccinia spp P. graminis, Pythium spp, Ramularia spp of which R betae , Rhizoctonia spp for example R. solani, Rhizopus spp for example R. nigricans, Rynchosporium spp as R. secalis, Sclerotinia spp as S sclerotiorum, Septoria spp for example S. nodorum or S.tritici, Sphaerotheca spp as S. macularis, Taphrina spp for example T pruni, Uncinula spp for example U. necator, Ustilago spp for example U. tritici and Venturia spp for example V. inaequalis.

A particular pathogen is Venturia inaequalis, which is responsible for apple tree mange.

Examples of bacteria that affect crops include mainly Corynebacterium, Clavibacter, Curtobacterium, Streptomyces, Pseudomonas, Xanthomonas, Erwinia spp and particularly E. amylovora, E. carotovora, E. chrysanthemi. Examples of viruses that affect crops are for example tobacco mosaic viruses, or potato virus Y.

A particular objective of the present application refers to the use of walls or bark of yeast (s) as an active material, for the treatment of mange, mainly in fruit trees, mainly apple tree mange. Another object of the invention relates to a method for the treatment of scabies, mainly in fruit trees, mainly apple trees, which comprises the application on the plant of walls or bark of yeast (s), or of a composition comprising walls or yeast bark (s).

The invention relates to the induction or stimulation of the natural defenses of a plant against scabies, mainly in fruit trees, which comprises the application on said plant or one of its parts of walls or bark of yeast (s), or of a composition comprising walls or crusts of yeast (s).

The invention relates to the prevention or braking of the development of Venturia strains resistant to a fungicidal agent, characterized in that the treatment (s) of the plant with said fungicidal agent is (are alternated (s)

or combined (s) with a treatment (s) of said plant with bark or walls of yeast (s), or a composition comprising bark or walls of yeast (s). In a specific way, the procedure is used to curb the development of resistant strains of Venturia inequalis and / or Venturia pirina.

The use of bark or walls of yeast (s), or of a composition comprising bark or walls of yeast (s), for the prevention or treatment of scabies in organic or organic farming is described.

The present application describes the use of walls or bark of yeast (s), or a composition comprising walls or bark of yeast (s), for the treatment of cercosporiosis, mainly in beet. Another object of the invention relates to a process for the treatment of cercosporiosis, mainly in beet, which comprises the application on the floor of walls or bark of yeast (s), or of a composition comprising bark or walls of yeast .

The invention describes a method for inducing or stimulating the natural defenses of a plant with cercosporiosis, mainly in beet, which comprises the application on said plant or one of its parts of walls or yeast bark (s), or of a composition comprising bark or walls of yeast (s).

Another object of the invention lies in the use of bark or yeast walls (s) as an active material, for the prevention or treatment of cercosporiosis in organic or organic farming.

Other aspects and advantages of the present invention will appear upon reading the following examples, which should be considered as illustrative and not limiting.

Examples

Example 1: efficacy test against apple tree scabies of compositions comprising yeast barks.

Scabies is the main disease of fruit trees with seeds. On the apple tree, the responsible agent is Venturia inaequalis.

In the absence of treatment, scabies would cause losses in yields and crop quality, reaching up to 70% of the value of the crop, which leads the arboricultor to protect his orchard at the price of 10 to 15 phytosanitary treatments throughout it's from the season. The economic and environmental consequences are very hard.

The quantities of active materials thus contributed represent by themselves more than half of the contributions of globally applied agrochemical active substances on apple trees, one of the most consumer crops of phytosanitary products. Such contributions generate significant amounts of waste in soils, waters, and on the same fruits.

Current treatments available against v can be regrouped into four product families:

Contact products, non-penetrating, and therefore sensitive to rain washing, non-mobile and therefore leaving unprotected organs after treatment, which means re-treating as soon as new organs have sprouted (leaves, fruits) or fattened.

Anilinopyrimidines, partially penetrating, but which do not protect the organs that appeared after treatment, and select resistant strains of Venturia.

Strobilurins, slightly mobile in the plant, and develop resistant strains.

Sterol synthesis inhibitors (IBS) are systemic: they spread through the sap. They protect the organs that develop after treatment. These products are also subject to resistance development.

Limiting resistance requires limiting the repetition of treatments based on agrochemicals of the same family to 2 or 3, and alternating families.

In this context, the provision of a new composition that acts differently, producing a lasting effect, and not providing chemical waste, would be a real advantage for the arboricultor, the consumer and the environment.

This first example demonstrates the effectiveness of bark and yeast walls for the protection of apple trees against scabies.

Material and methods

In trial it is carried out under greenhouse under controlled conditions, on young apple trees from seedlings.

A population of seeds coming from an open pollination at 4 ° C in petri dishes filled with sand and moistened to saturation for 90 to 120 days is germinated.

The wetting of the sand is done every 15 days.

From the germination of the first pips, a sowing of all the pips is carried out in plastic terrines of 40 x 30 x 15 cm, at the rate of sixty pips per terrine, of which only 50 will be retained for the test. The seeds are placed in rehydrated peat tablets on a bed of mulch (CombiTree B MG of the DCM brand) containing a mixture of fertilizers (NPK 7-7-10 at 2 kg / m3 and NPK 15-8-12 diffusion slow, at 2 kg / m3).

The terrines are covered with mulch, moistened and placed at a temperature close to 10 ° C for a week before putting them in a greenhouse at 18 ° C. If necessary, an insecticidal protection is ensured during the test.

Seedlings are treated from the stage 3-4 unfolded leaves, at the rate of two successive treatments, separated by a week. The pathogen (Venturia inaequalis) is inoculated three days after the second treatment. The annotations are made 14 days and 21 days after inoculation.

The statistical device is 3 repetitions, each corresponding to a terrine of 50 seedlings.

The test compares 3 doses of an aqueous suspension (demineralized water) OY of yeast barks that respectively provide 2.5 mg / l, 25 mg / l and 250 mg / l of bark) to a control treated with demineralized water.

The barks used here correspond to the product Springcell 8001 of Biospringer SAS, (Maisons-Alfort, France), composed of crusts with 96% dry matter.

Table 1

Objects

products Concentrations (mg / l)

OY1

Bark 250

OY2

Bark 25

OY3

Bark 2.5

Untreated witness

Water -

The treatment is done by spraying after stirring, with a 500 ml manual sprayer (BIRCHMEIER brand). Spraying at the runoff limit is interrupted. The last sheet displayed on the day

preceding the treatment is called "F1", and is located by a loop attached to the petiole.

The newly formed or unfolded sheet after treatment, and which has not received the composition of

treatment, is called "F0". When this sheet forms but does not unfold during treatment, it is

masks during spraying. When inoculation occurs 3 days after treatment, F0 is at least partially deployed during this operation.

The inoculum of Venturia inaequalis is prepared as follows.

Mangy leaves have been collected during the summer in different orchards. After drying for 20 days, the leaves are put in plastic bags and stored in the freezer at -18 ° C. On the day of use, these leaves are placed in a 1-liter bottle, which contains 200 ml of rainwater, then shaken manually for 10 minutes. The suspension is filtered with ethamine, and the volume obtained is measured.

The conidia are counted under a microscope on a Bürker hematocimeter, at the rate of 2 counts of 2x144 square, making the average. This number of conidia is multiplied by the Bürker constant (250000), and, to obtain a number of viable conidia, corrected with a germination coefficient of the conidia, the same result of a test carried out the previous day.

On this basis, the spore suspension is diluted to obtain 150,000 viable conidia per ml. It takes then approximately 1 liter of suspension to inoculate 1000 seedlings.

The seedlings are inoculated by manual spraying, with a suspension of 150,000 viable conidia of Venturia per ml, and transferred to a humid saturation chamber for 48 hours.

The annotations refer to F1 and F0. They express the sporulant leaf surface, as a percentage of the leaf surface.

The average of the 50 seedlings is calculated for each repetition. The average of the 3 repetitions of each object (or modality) leads to the average per object. Finally, the efficiency according to Abott is calculated, according to the formula:

Efficacy = [(Annotation "water") - (Annotation object tested)] / (Annotation "water")

Results

The results are presented in table 2 below. Table 2

Notations

Sporulant surface / sheet (%) Efficiency (%)

Objects

F1 F0 F1 F0

Water

41.46 15.83 0 0

OY1

17.75 10.39 57.2 34.4

OY2

14.59 7.6 64.8 52.0

OY3

16.9 8.9 59.2 43.8

The results obtained show that the products according to the invention are capable of inducing a significant decrease in the sporulant surface, both on treated F1 sheets, and on the F0 sheets formed after the treatment.

In the latter case, since the walls or barks cannot penetrate the plant to be vehiculized by the sap, it is probably not a system. On the contrary, this result suggests an inducing effect of the plant's natural defense mechanism.

Example 2: efficacy test against scabies of compositions comprising yeast barks.

The test was carried out on grafted seedlings, in pots. Three varieties were used: "Reinette des Capucins", "Jonagold", and "Reinette de Waleffe", grafted onto an M9 graft holder. The growth of varieties 15 under a plastic tunnel shifts so that the seedlings were all in the same stage during the test. Reinette de Waleffe, then 15 days later, Reinette des Capucins, then 1 week later, Jonagold.

The seedlings were labeled with the name of the variety and the number of the treatment they will receive before inoculation.

The methods of treatment, inoculum preparation, inoculation and annotation are similar to those described in example 1.

The seedlings were treated at the rate of two successive treatments, separated by 10 days. The day after

second treatment, the last leaf deployed "f1" is located.

Two days after the second treatment, the pathogen (Venturia inaequalis) is inoculated at the dose of 1.5 * 105 conidia / ml, directly in a humid chamber. The inoculation is followed by a 48-hour incubation in a humid chamber at 18 ° C, then the seedlings are kept in the cubicles conditioned at 18 ° ± 2 ° C and 80 ± 10% of H.R.

The device studied is as follows (table 3):

Table 3

Varieties

 Objects products Concentrations (mg / l)

Jonagold

OY2 Bark 25

Jonagold

SLIME 1-aminobutyric acid 2000

Jonagold

Witness Water -

Waleffe

OY2 Bark 25

Waleffe

SLIME 1-aminobutyric acid 2000

Waleffe

Witness Water -

Varieties

 Objects products Concentrations (mg / l)

Capucins

OY2 Bark 25

Capucins

SLIME 1-aminobutyric acid 2000

Capucins

Witness Water -

The OY2 yeast barks correspond to an aqueous suspension at 25 mg / l of the Springcell 8001 product of Biospringer SAS, (Maisons-Alfort), France), which consists of 96% dry matter barks.

The annotations were made on the twenty-first day after inoculation on the last 2 treated leaves (F2 and F1) and the neoformed leaf after the F0 treatment, on approximately 25 shoots per object. The set of results is shown below, table 4.

Table 4

Sporulant surface / sheet (%)

Efficiency (%)

Varieties

Objects F2 F1 F0 F2 F1 F0 H treated All H

Capucins

Water 17.5 19.8 7.8 - - - - -

Capucins

Slime 4.2 8.0 4.3 75.8 59.8 45.3 67.8 60.3

Capucins

OY2 3.2 7.1 4.2 81.6 64.4 46.7 73.0 64.2

Jonagold

Water 40.3 36.1 22.4 - - - - -

Jonagold

Slime 6.0 17.9 13.9 85.1 50.5 38.0 67.8 57.9

Jonagold

OY2 13.6 12.7 13.7 66.2 64.8 38.8 65.5 56.6

Waleffe

Water 31.1 31.4 23.3 - - - - -

Waleffe

Slime 18.6 20.4 20.9 40.3 35.2 10.3 37.8 28.6

Waleffe

OY2 5.5 14.5 10.7 82.2 54.0 54.1 68.1 63.4

All

Water 29.6 29.1 17.8 - - - - -

All

Slime 9.6 15.4 13.0 67.6 47.1 27.0 57.4 47.2

All

OY2 7.5 11.4 9.5 74.8 60.8 46.6 67.8 60.8

All varieties and all leaves confused, the effectiveness is average: 60.8%. On average it is 68.8% on 10 of the F2 and F1 treated sheets, and 46.6% on the neoformed F0 sheets.

This efficiency is close, but always superior to that obtained with the BABA in the same period.

An annotation at 41 days after treatment gives the following results:

Table 5

Varieties

Objects F2 F1 F0 F-1 F-2 Treated sheets Neoformed leaves All the leaves

Capucins

Water 90.00 33.55 36.13 22.57 4.33 - - -

Capucins

Slime 4.00 10.62 14.07 9.95 4.00 88.2 55.5 77.1

Capucins

OY2 1.80 7.76 15.69 11.07 1.92 92.3 54.5 79.5

Jonagold

Water 50.73 67.69 64.92 51.54 22.44 - - -

Jonagold

Slime 10.57 27.57 37.79 29.57 3.43 67.8 49.0 57.7

Jonagold

OY2 20.42 35.71 52.19 32.56 12.29 52.6 30.1 40.5

Waleffe

Water 50.00 54.30 61.35 56.06 20.18 - - -

Waleffe

Slime 15.55 42.00 41.05 45.16 18.42 44.8 24.0 33.0

Waleffe

OY2 0.50 15.40 18.59 17.31 8.07 84.8 68.0 75.2

All

Water 63.6 51.8 54.1 43.4 15.7 - - -

All

Slime 10.1 26.7 31.0 28.2 8.6 68.1 40.1 54.2

All

OY2 7.6 19.6 28.8 20.3 7.4 76.4 50.0 63.4

On this date, new leaves were deployed: F-1 and F-2, and the disease progressed by natural contamination, without a new inoculation.

5 All varieties and all leaves confused, the effectiveness is average: 63.4%. On average it is 76% on the treated leaves, F2 and F1, and 50% on the neoformed leaves: F0, F-1 and F-2.

Again, this efficiency is close, but always superior to that obtained with the BABA in the same period.

The very long persistence of the fungicidal effect, much higher than that of agrochemicals (of the order of 7 to 15 days) will be noted.

10 Example 3: protection test against beet cercosporiosis

A small-scale greenhouse trial was carried out for beet cercosporiosis. This fungal disease is caused by Cercospora beticola.

Grains were sown in the FORTIS beet variety recognized as sensitive to cercosporiosis in germinators. Upon leaving, the young seedlings were planted in pots kept in a greenhouse at 24 ° C, under a

15 photoperiod of 16 hours of light per day. The experimental device comprised 2 blocks (2 repetitions) of 8 plants per modality (or object).

The seedlings were treated only once, in the 4-leaf stage by spraying on the leaves of preparations containing yeast barks.

The preparations, encoded OY, were composed of yeast barks (Springcell 8001 of Biospringer 20 SAS, composed of 96% dry matter barks) in aqueous suspension.

The treatment consisted of spraying the preparations obtained, with the help of a manual sprayer, after stirring. The treatment, carried out in the 4-leaf beet stage, pointed to the first 2 fully developed leaves and ensured coverage of the 2 leaf faces up to the runoff limit.

5 The treated plants were kept in an enclosure under an atmosphere kept humid by regular watering of the substrate.

The seedlings were inoculated 1 week later, in the 6-leaf stage, by spraying a conidial suspension of C. beticola strain 524, at 20,000 spores per ml, on the two leaf faces, to the runoff limit, then the Plants were kept under a humid atmosphere.

The 524 strain of Cercospora beticola provided by the Phytopathology Unit of the University Faculty of

10 Gembloux (Belgium) was chosen because of its aggressiveness. The strain was grown in Petri dishes on V8 medium in the form of individualized colonies. After 5 days of incubation in the dark in a culture chamber at a temperature of 24 ° C, the individualized colonies were recovered in a sterile tube containing 3 ml of sterile distilled water. The preparation thus obtained was then stirred with vortex to release the conidia of the pathogen. The conidia suspension was then used to sow new Petri dishes containing freshly prepared V8 medium. Crops

15 so prepared were incubated at 24 ° C in a culture chamber where the photoperiod was 16 hours of light. After one week of incubation, the culture was recovered and a conidia suspension was prepared by surface scratching of the culture in the distilled water using a Scalpel slat. The count of conidia in the conidia suspension was then counted using the Bürker cell, and the suspension was adjusted to 20,000 conidia per milliliter in distilled water.

20 The plants so inoculated were kept in these conditions of strong humidity.

The importance of the symptoms was evaluated one month later with the help of the visual scale used by the Royal Belgian Institute for Beet Improvement (IRBAB), which comprises 10 values from 0 to 9, on which the value 9 corresponds to 100% of healthy leaf surface (not covered by lesions), and the value 0 to 0% of healthy leaf surface (not covered by lesions).

The treatment was carried out by means of a suspension of barks encoded OY, with different concentrations, OY1, OY2, OY3, described in Table 6 below.

Table 6

Objects

products Concentrations (mg / l)

OY1

Bark 250

OY2

Bark 25

OY3

Bark 2.5

Untreated witness

Water -

The grade obtained for each object, in the middle of two repetitions is collected in table 7 below. 30 Table 7

Objects

Average grade Healthy surface in% IRBAB qualifier

Untreated witness

3.94 85 Insufficient

OY1

4.69 91 Acceptable

OY2

6.34 97.5 Very acceptable

OY3

6.22 97.1 Very acceptable

In this example 3, the cortices allow to go from an average appreciation of damage classified as insufficient to a qualified appreciation as very acceptable.

Example 4: Wheat protection test against septoriosis

Septoriosis (Septoria nodorum, and / or Septoria tritici) is the main foliar disease of wheat in Europe, still causing yield losses that can reach up to 40% of the crop. The chemical fight generally consists of a systematic treatment in the stage 2-3 knots that can be followed by a

5 spike stage treatment.

This test shows that the early contribution of a preparation based on walls or bark of yeasts allows to delay the onset of the disease, and replace the first chemical treatment. Which translates into benefits on the toxicological plans for the operator and the consumer (waste) and on the environmental plan.

Material and methods

10 The test was carried out in France, in the countryside, on a winter soft wheat crop of the Orvantis variety, planted on 06.10.2005.

The test was conducted according to method CEB No. M189 (Biological Testing Commission, of the French Association for Plant Protection, Paris), and respecting Good Experimental Practices.

The statistical device consists of randomized Fisher blocks. Each modality has 4 repetitions, 15 corresponding each to an elementary plot of 8x 2.5 cm (20 m2).

The test consists of comparing the effect of the yeast bark used in aqueous suspension (Springcell 8001 from Biospringer SAS, composed of 96% dry matter bark), encoded OY, when used at the beginning of a septoriasis treatment program .

The chemical reference used here is Opus (epoxiconazole 125 g / L, BASF Agro) used at 1 L / ha.

20 The treatment was carried out at a rate of 200 L / ha with the help of a spray truck equipped with a spray ramp of 2.50 m.

Following the modalities, the treatment program varies as indicated in the following table. All modalities receive Opus at 1L / h 40 days after the first treatment.

Table 8

No. of modality

T at the stage “end of macollación / tang at 1 cm” T + 7 days Stadium "2-3 knots" 2nd treatment at T + 40 days

one

Dry untreated witness - - OPUS 1 L / ha

2

Witness treated with water - - OPUS 1 L / ha

3

- - Opus 1 L / ha OPUS 1 L / ha

4

OY at 2.5 g / ha - - OPUS 1 L / ha

5

OY at 25 g / ha - - OPUS 1 L / ha

6

OY at 250 g / ha - - OPUS 1 L / ha

7

OY at 50 g / ha - - OPUS 1 L / ha

8

OY at 25 g / ha OY at 25 g / ha - OPUS 1 L / ha

Five annotations were made on the trial period in order to evaluate the frequency and intensity of septoriosis attack:

-

Annotation 1: 04.04.06 to T1 (BBCH 31) spike stage at 1 cm.

-

Annotation 2: 28.04.06 to T3 (BBCH 32) second knot stage; 30 -Annotation 3: 15.05.06 to T4 (BBCH 39) stadium last leaf deployed.

-

Annotation 4: on 29.05.06 at T4 + 15 days (BBCH 55) semi-spike stadium

-Annotation 5: 22.06.06 at BBCH 72 stadium, aqueous grain. The timing of the trial is therefore the following (N = annotation):

For each score, the frequency and intensity were estimated on 3 foliar stages (F1, F2, F3), 5 designating F1 as the last fully developed visible leaf, on 25 randomly selected feet. The frequency corresponds to a percentage of leaves affected by septoriosis.

The intensity corresponds to an average percentage of leaf area reached by the disease. The average of the 25 feet is calculated for each repetition. The average of the 4 repetitions for each modality leads to the average by modality. Finally, efficiency is calculated according to Abott.

10 Results During the first 4 entries, the intensity of the disease remained low (<3% in the untreated control).

The effectiveness of OY modalities does not appear statistically significant. In the 5th entry, the disease is declares and intensity greater than 95% in the untreated control on sheet F3. On this date, the results, significance of the differences, and percentages of effectiveness are as follows:

15 Table 9

% affected leaf area

F1 sheet =% efficiency F2 sheet =% efficiency Med F1 effectiveness + F2 =%

1 dry dry

13.5 A - 45.2 A - 29.35 TO -

2.Testigo treated with water

12.7 A - 33.6 B - 23.15 TO -

3.Opus 1 L / ha

3.4 B 74.80% 13 C 71.20% 8.2 B 73.00%

4.OY at 2.5 g / ha

4.4 B 67.40% 13.3 C 70.60% 8.85 B 69.00%

5.OY at 25 g / ha

4.7 B 65.20% 15.3 C 66.10% 10 B 65.65%

6.OY at 250 g / ha

3.7 B 72.60% 13.9 C 69.20% 8.8 B 70.90%

7.OY at 50 g / ha

3.2 B 76.30% 15.4 C 65.90% 9.3 B 71.10%

8.OY at 25 g / ha + OY at 25 g / ha

3.6 B 73.30% 9.7 C 78.50% 6.65 B 75.90%

F3 sheet =% efficiency

96.3 AB -

96.1 AB -

87.5 AB 9.10%

98.2 to -2.00%

95.6 AB 0.70%

84.8 AB 11.90%

92.9 AB 3.50%

90.2 AB 6.30%

The results show a significant decrease in the intensity of the disease on F1 and F2, 80 days after application in the first treatment, and whatever the tested dose of OY. Better yet, the efficiencies obtained on average over F1 and F2 with OY are all statistically equivalent to that obtained with the

20 chemical reference Opus.

Example 5: Protein pea protection test against anthracnose.

Anthracnose is one of the main foliar diseases of the pea and strongly penalizes grain yield. It is caused by an Ascochyta pisi mushroom.

Material and methods

The test was carried out in France, in the countryside, on a spring protein pea crop of the Lumina variety. The crop was sown on March 22. The test was carried out according to method CEB No. M215 (Biological Testing Commission of the Association

5 French for Plant Protection, Paris), and respecting Good Experimental Practices).

The statistical device consists of randomized Fisher blocks. Each modality has 4 repetitions, each one corresponding to an elementary plot of 8x 2.5 m (20 m2). The essay includes:

-

 2 controls: 1 dry and one treated with water: modalities 1 and 2 10 - 1 chemical reference (Dithane Neotec: Mancozeb 75%, Dow Agroscience) applied twice: modality 3

-

 3 modalities of yeast barks, used in aqueous suspension at different concentrations (Springcell 8001 of Biospringer SAS, composed of 96% dry matter barks), OY encoded, modalities 4 to 6:

○ Modality 4: 25 g / ha

○ Modality 5: 250 g / ha 15 ○ Modality 6: two treatments spaced for one week, at 25 g / ha then at 25 g / ha

The treatment was carried out at a rate of 200 L / ha with the help of a spray truck equipped with a ramp of 2.50 m. The first applications were made on 05.17.2006 in the 7-8 leaf stadium. Artificial contamination was carried out on 19.05.06 with the help of mycelium and fresh spores of Ascochyta pisi on 20 barley grains (20 kg of grains / 1000 m2), provided by the company ARBIOTECH. The positioning of the treatments is as follows: Table 10

No. of modality

T: 1st leaf trifoliate T + 7 days Flowering

one

Witness - -

2

Water witness - -

3

Dithane Neotec (2.1 kg / ha) - Dithane Neotec (2.1 kg / ha)

4

OY at 25 g / ha - -

5

OY at 250 g / ha - -

6

OY at 25 g / ha OY at 25 g / ha -

The annotation was made on 10.06.06 in the BBCH 67 stage (flowering stage), in order to evaluate the frequency and intensity of the anthracnose attack on the leaves.

The timing of the test is therefore the following:

During the annotation, the frequency and intensity were estimated over 3 foliar stages (low-medium-high) over 25 feet per elementary plot. The frequency corresponds to a percentage of leaves affected by the disease.

5 The intensity corresponds to an average percentage of leaf area reached. The average of the 25 feet is calculated for each repetition. The average of the 4 repetitions for each modality leads to the average per modality. Finally, efficiency is calculated according to Abott.

An analysis of variance was performed, as well as a Neuman and Keuls trial, in order to assess the significance of the differences between modalities (same letters: results not different to the risk of 5%; different letters: 10 results different from the risk of 5%).

Results Despite artificial contamination, anthracnose infections were quite low (intensity < 8% in the untreated witness).

In the annotation, the results and the percentages of effectiveness are as follows. 15 Table 11: Intensity of anthracnose attacks on leaves and efficacy

Intensity of anthracnose attacks on leaves in% (06.10.2006 BBCH 67) and effectiveness (Ef)

No.

Treatments Low stage Ef Middle stage  Ef Medium low + medium High stadium

one

Dry witness 6.20 A - 3.35 A - - - 0.07 B

2

Witness treated with water 5.83 AB - 3.31 A - - 0.2 B

3

Dithane Neotec (2.1 kg / ha) (2 times) 4.35 C 30% 2.07 BC 38% 3. 4% 0 B

4

OY at 25 g / ha 4.72 C 24% 2.25 BC 33% 29% 0.038 A

5

OY at 250 g / ha 4.87 CB twenty-one% 1.52 C 54% 38% 0.02 B

6

OY at 25 g / ha + OY at 25 g / ha 4.33 C 30% 2.45 B 27% 29% 0.07 B

Since the upper part is not attacked yet, the evaluation is done in the lower and middle part. All tested doses of OY show an efficacy equivalent to that of chemical reference.

Example 6: protection test of the vineyard against the oidium (Ersiphe necátor).

The oidium (Uncinula necator, Erysiphe necator) is a fungal disease of the vineyard present in all vineyards with different intensities according to regions and strains. It is the most widespread vineyard disease in the world. The ocean attacks all the organs of the vineyard and can cause very important production losses.

5 Material and methods

The test was carried out under greenhouse, in pots, on young plants of the Cinsaut strain. The conditions were controlled, and the artificial contamination.

The seedlings were produced in the greenhouse from cuttings of one eye, and homogeneous batches were formed.

The statistical device comprised 6 repetitions of a pot each. The repetition that gave the result 10 furthest from the average was eliminated during the analysis.

The treatments were carried out with a spray projection jet bank and constant pressure on the whole plant, and the plants of the same batch were treated simultaneously. The bank consists of a spray car, which travels on a rail with constant speed and comprises 5 mouths (2 on each side and one above the vegetable). The treatment volume was equivalent to 600 liters per ha.

15 During the treatment, the leaf levels were marked with a fixed color loop under the 3rd leaf unfolded so that the effect of the treatment on the neoformed leaves (formed after the treatment) can be observed.

The treatments were applied 14 and 7 days before the artificial contamination. A batch of plants was treated twice, that is 14 days after 7 days before contamination.

The products tested were yeast barks used in aqueous suspension (Springcell 8001 of 20 Biospringer SAS, composed of barks of 96% dry matter), OY-encoded.

4 doses were tested:

-

Dose N / 10: 2.5 g / ha

-

Dose N: 25 g / ha

-

Dose 2N: 50 g / ha 25 -Dose 10N: 250 g / ha

The doses were tested following the following schedule: Table 12

Number of days between treatment and inoculation

No. of modality

T 14 d before inoculation T 7 d before inoculation

one

Untreated witness -

2

OY N (25 g / ha) -

3

- Untreated witness

4

- OY N / 10 (2.5 g / ha)

5

- OY N (25 g / ha)

6

- OY 2N (50 g / ha)

7

- OY 10N (250 g / ha)

8

OY N (25 g / ha) OY N (25 g / ha)

The fungal material consisted of a set of conidia from a strain of normal sensitivity to fungicides.

All the test plants were contaminated by dry sprinkling of spores above the plants located inside a plexiglass inoculation tower.

The fungal material used is constituted by a set of spores of an oid medium strain multiplied previously in large numbers on surviving leaves or on plants. The inoculum used has 12 to 14 days for the oídium of the vineyard. The inoculum quality was verified by means of a Malassez cell located at the level of the plants inside the inoculation tower. A density of 800 to 1000 spores per cm2 was used.

All plants were incubated after contamination in an acclimatized room at 21 ± 2 ° C and illuminated 14 hours per day. Each test condition is perfectly isolated from the others in small enclosures.

The plants were kept in these conditions for 14 days. At the end of this period, an annotation of fungal damage was made.

Annotations

The leaves of the upper foliar levels F2, F1 and F0 (level formed after treatment) were indicated. Each sheet is designated for a visual observation. The frequency (percentage of leaves affected by the disease) is not indicated since it is equal to 100. The intensity (average percentage of leaf area affected) was evaluated according to a scale of 1 to 100.

The efficiency according to Abott is calculated on the average intensities, and an analysis of variance is carried out. An essay by Neuman and Keuls allows us to evaluate the significance of the differences between modalities (same letters: results not different from the risk of 5%; different letters: results different from the risk of 5%).

Results

Table 13: damage rate on plants treated on different dates (14 and 7 days before inoculation by E. necator) with the OY coded specialty (Springcell 8001 from Biospringer SAS). Observation on sensitive sheets F2, F1, F0:

Modality

Number of days between treatment and infection Attack Intensity (%) Effectiveness

Witness

14 days 47.4 AB -

N

14 days 48.0 AB 0

Witness

7 days 61.3 A -

N

7 days 40.1 AB 34.6

N / 10

7 days 32.7 ABC 46.7

2N

7 days 23.3 BC 62

10N

7 days 21.3 BC 65.3

N + N

14 days + 7 days 14.1 C 70.3

A single application 14 days before inoculation has not allowed to protect the leaves.

Conversely, applications made 7 days before contamination produce a protective effect against oidium The strongest doses (2N, 50 g / ha and 10N, that is 250 g / ha) have a more significant effect. An application of 25 g / ha (dose N) 14 days before inoculation, followed by an identical one, 7 days before

inoculation, gives an excellent result. Example 7: protection test of the vineyard against mildew (Plasmopora viticola). The mildew is due to a fungus (Plasmopora viticola). Present in varying degrees in most vineyards of the

world, damages the yield and quality of the crop, to the point of being able to annihilate it in the absence of treatment.

Material and methods

The test was conducted under greenhouse, in pots, on young plants of the Cabernet-Sauvignon strain. The conditions were controlled, and the artificial contamination.

The seedlings were produced in the greenhouse from cuttings of one eye, and homogeneous batches were formed.

5 The statistical device comprised 6 repetitions of 1 pot each. The repetition that gave the result furthest from the mean was eliminated during the analysis.

The treatments were carried out with a spray bank of projected jet and constant pressure on the whole plant, and the plants of the same batch were treated simultaneously. The bank consists of a spray car, which travels on a rail with constant speed and comprises 5 mouths (2 on each side

10 and one above the vegetable). The treatment volume was equivalent to 600 liters per ha.

During the treatment, the foliar levels were marked by a fixed color loop under the 3rd leaf deployed so that the effect of the treatment on the neoformed leaves (formed after the treatment) can be observed.

The treatments were applied 14 and 7 days before the artificial contamination. A batch of plants was treated twice, that is 14 days after 7 days before contamination.

15 The products tested were yeast barks used in aqueous suspension (Springcell 8001 from Biospringer SAS, composed of 96% dry matter barks), OY-encoded.

4 doses were tested:

-

Dose N / 10: 2.5 g / ha

-

Dose N: 25 g / ha 20 -Dose 2N: 50 g / ha

-

10N dose: 250 g / ha

The doses were tested following the following schedule: Table 14

Number of days between treatment and inoculation

No. of modality

T 14 d before inoculation T 7 d before inoculation

one

- -

2

- OY N / 10 (2.5 g / ha)

3

- OY N (25 g / ha)

4

- OY 10N (250 g / ha)

5

- OY 2N (50 g / ha)

6

OY N (25 g / ha) OY N (25 g / ha)

7

OY N (25 g / ha) -

The plants were inoculated simultaneously by spraying a suspension of sporophytes from a strain of Plasmopora viticola normally sensitive to fungicides.

The sporophyte suspension was prepared just before contamination. The sporulations of the fungus were recovered by washing infected leaves with the permutated water. An assessment was performed with the help of a Malassez cell. The concentration used was 50,000 spores / ml.

30 10 ml of spore suspension per plant was sprayed on the underside of the leaves. Each plant was contaminated individually, at all levels of leaves present.

The plants were then regrouped by modality, and the modalities were isolated from each other by enclosures

closed. The plants were then kept under nebulization to favor the development of the disease, at a temperature of 21 ° C, and 14 hours of lighting per day, for 8 days. At the end of this duration, they were made

5 the annotations. Annotations Each sheet is marked for visual observation. The frequency (percentage of leaves affected by the disease) is not

noted since it is equal to 100.

Intensity (mean percentage of leaf area affected) was evaluated on a scale of 1 to 100. 10 Efficacy according to Abott was calculated over the mean intensities.

Results

Intensity results and efficiencies were regrouped depending on whether they were treated sheets or leaves

neoformed, therefore untreated. Table 15

Treated sheets

Neoformed leaves

Modality

Number of days between treatment and infection Attack Intensity (%) Efficiency (%) Attack Intensity (%) Efficiency (%)

Witness

14 days 68.0 - 67.3 -

N

14 days 31.3 54 45.0 33.1

Witness

7 days 82.0 - 48.1 -

N / 10

7 days 62.7 23.5 40.5 15.8

N

7 days 54.7 33.3 23.6 50.9

2N

7 days 50.0 39 23.5 51.1

10N

7 days 42.7 47.9 28.0 41.8

N + N

14 days + 7 days 25.4 62.6 40.7 39.5

All the doses of barks tested had an efficacy on the mildew of the treated or neoformed leaves, whatever the period separating the treatment from contamination. The effectiveness tends to grow with the dose. Example 8: test in full field of protection of the vineyard against the mildew. Material and methods 20 The test was carried out in France, near Bordeaux, in the countryside, on the Cabernet-Sauvignon vineyard. The essay was carried out respecting Good Experimental Practices. The installation of test plots and the first applications were carried out on 05.25.2006 at the BBCH stadium

55. The statistical device consists of randomized Fisher blocks. Each modality has 4 repetitions, each corresponding to an elementary plot of 10 strains. The essay includes:

25 -1 untreated witness;

-

2 modalities OY in aqueous suspension (Springcell 8001 from Biospringer SAS, composed of crusts with 96% dry matter).

The treatment was carried at a rate of 1000 L / ha with the help of a Solo brand atomizer. The positioning of the treatments consists of a weekly application from May 25 (BBCH55 stage).

Table 16

Modality Nº

Weekly treatment

one

Untreated

2

OY 4N (100 g / ha)

3

OY 10N (250 g / ha)

The annotation was made before application. When the disease was late, the annotation was made on 18.08.2006. The frequency and intensity on the leaves were estimated on 50 leaves taken at random, per elementary plot, it is

5 say 5 leaves per foot of vineyard. The frequency corresponds to a percentage of leaves affected by the disease. The intensity corresponds to an average percentage of leaf area affected. The average of the 50 sheets was calculated for each repetition. The average of the 4 repetitions for each modality

led to the average by modality.

10 Finally, the effectiveness according to Abott was calculated over the average intensities. Results The results and the percentages of effectiveness are as follows.

Table 17: Mildiú annotation on leaves of August 18 - BBCH 83

No.

Treatments Frequency / sheets (%) Intensity / sheets Efficiency (%)

one

Untreated witness 48.8 13.3 -

2

OY4N (100g / ha) 53.0 9.9 25.56

3

OY10N (250g / ha) 45.5 5.5 58.64

15 Product efficacy was observed at the 2 doses tested, ie 100 and 250 g / ha.

Claims (24)

one.

Use of walls or bark of yeast (s) as active material, for the treatment or protection of plants against diseases caused by pathogens or for the induction or stimulation in a plant of natural defenses against pathogens.

2.

Use according to claim 1, characterized in that the plant is chosen from grasses and dicotyledons, annual, bi-annual and perennial plants, vegetables, oilseeds, protein crops, potatoes, beets, sugar cane, tobacco, woody plants, trees, fruit or not, vines and vegetables for decoration.

3.

Use according to claim 1 or 2, characterized in that the plant is a fruit tree.

Four.

Use according to any one of the preceding claims, characterized in that the pathogen is a fungus, a virus, a bacterium, a mycoplasma, a spiroplasma or a viroid.

5.

Use according to claim 4, characterized in that the pathogen is chosen among the fungi of the genera Alternaria spp, Ascochyta spp, Botrytis spp, Bremia spp, Cercospora spp, Cladosporium spp, Colletotrichum spp, Erysiphe spp, Fusarium spp, Glocosporium spp, Guignardia spp, Helminthosporium spp, Marssonina spp, Monilia spp, Mycosphaerella spp, Penicilium spp, Peronospora spp, Pezicula spp Phragmidium spp, Phytophtora spp, Plasmopara spp, Podosphaera spp, Pseudocercosporella spp, Pseudocerpppppppp, Pysepppc spp, Pseudocerpper spp, Pseudocercosprella spp, Pseudocerpper spp, Pseudocerpper spp, Pseudocerpper spp, Pseudocerptos spp, Pseudocercosporella spp Ramularia spp, Rhizoctonia spp, Rhizopus spp, Rynchosporium spp, Sclerotinia spp, Septoria spp, Sphaerotheca spp, Taphrina spp, Uncinula spp, Ustilago spp and Venturia spp.

6.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeast (s) are of the genus Saccharomyces.

7.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeast are susceptible to be obtained by lysis of yeast cells, separation of the soluble and insoluble parts, then recovery of the insoluble part.

8.

Use according to claim 7, characterized in that the insoluble part is recovered by elimination of the soluble part by centrifugation.

9.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are delipidated.

10.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are further associated with one or more formulation agents.

eleven.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are further associated with a fungicidal, antiviral or antibacterial agent.

12.

Use according to claim 11, wherein the fungicidal agent is chosen from organic agrochemical fungicides or inorganic mineral fungicides based on sulfur and / or copper.

13.

Use according to claim 12, in which the organic agrochemical fungicidal agents are chosen from chloronitriles, carbamates, phthalimides, sulfa drugs, guanidines, quinones, quinoleins, thiadiazines, anilides, hydroxyanilides, phenylamides, imidazolinones, oxazolidinediones, strobilurines, cyanozimines, cyanozimines, cyanozimines, cyanozimines, cyanozimines, dicyanozines, cyanozimines, cyanozimines, cyanozimines, dicyanozines, cyanoimidaminines, cyanozimidines, cyanozimidines, cyanoimidaminines, cyanoimidaminines, cyanoimidaminines, cyanoimidaminines, cyanoimidaminines, cyanozimidines, cyanoimidaminines, cyanoimidaminines, cyanoimides , sitiofam, dicarboximides, fludioxonil, organophosphates, propamocarb HCl, diphenylamine, piridilaminas, inhibitors of sterol biosynthesis (IBS), imidazoles, pyrimidines, hydroxypyrimidines, anilinopyrimidines, triazoles, spiroxamine, morpholines and piperidines, fenhexamid, hymexazol, zoxamide, diethofencarb, benzimidazoles, pencicuron, quinoxifene, iprovalicarb, cymoxanil, dimetomorph, phosphonates, and triazines.

14.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are further associated with one or more eliciters of defenses compounds in the plants chosen from b-aminobutyric acid, 2,6-dichloroisonicotinic acid, acibenzolar-s -methyl, or certain algae extracts.

fifteen.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are in a concentrated phytosanitary composition, of dry or liquid formulation.

16.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are a composition ready for use.

17.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are administered by foliar or soil spraying.

18.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are administered in the roots by spraying in the soil, mechanical incorporation, in admixture with fertilizers, fertilizers, in premix, or others.

19.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are administered over all or a part of the plant, chosen from the leaves, stems, flowers, fruits, trunk or roots.

twenty.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are applied or used at an effective dose greater than 1 mg / l of bark or yeast walls when the product is sprayed to the limit of runoff, or greater than 1 g / ha in the case of spraying with low volume of water.

twenty-one.

Use according to the preceding claim, characterized in that the walls or bark of yeasts are applied or used at an effective dose comprised between 1 and 250 mg / l of bark or walls of yeasts when the product is sprayed to the limit of runoff , or between 1 and 250 g / ha in the case of spraying with low volume of water.

22

Use according to any one of the preceding claims, characterized in that the walls or bark of yeasts are used alternately and / or in combination with one or more other treatments.

2. 3.

Use according to claim 1, to prevent or curb the development of pathogens resistant to a family of active substances, characterized in that the plant is treated by the bark or walls of yeast (s) with a view to reducing strain selection pressure resistant to said family of active substances, or because the treatment (s) of the plant with a substance of said family of active substances alternates or combines with a treatment (s) of said plant with crusts or walls of yeast (s).

24.

Use according to any one of the preceding claims, characterized in that the walls or bark of yeast (s) derive from Saccharomyces cerevisiae.